Analysis of E. coli K5 capsular polysaccharide heparosan

作者：Ly Mellisa; Wang Zhenyu; Laremore Tatiana N; Zhang Fuming; Zhong Weihong; Pu Dennis; Zagorevski Dmitri V; Dordick Jonathan S; Linhardt Robert J^*

来源：Analytical and Bioanalytical Chemistry, 2011, 399(2): 737-745.

DOI：10.1007/s00216-010-3679-7

• E. coli K5 capsular polysaccharide
• Heparosan
• Molecular weight determination
• PAGE
• ESI-FT-MS
• Continuous-elution preparative electrophoresis

Heparosan is the key precursor for the preparation of bioengineered heparin, a potential replacement for porcine intestinal heparin, an important anticoagulant drug. The molecular weight (MW) distribution of heparosan produced by the fermentation of E. coli K5 was investigated. Large-slab isocratic and mini-slab gradient polyacrylamide gel electrophoresis (PAGE) were used to analyze the MW and polydispersity of heparosan. A preparative method that allowed fractionation by continuous-elution PAGE was used to obtain heparosan MW standards. The MWs of the heparosan standards were determined by electrospray ionization Fourier-transform mass spectrometry (ESI-FT-MS). A ladder of the standards was then used to determine the MW properties of polydisperse heparosan samples. Unbleached and bleached heparosan produced by fermentation of E. coli K5 had similar number-averaged MWs (M(N)), weight-averaged MWs (M(W)), and MW ranges of 3,000 to 150,000 Da.

• 出版日期2011-1
• 单位浙江工业大学